Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira

Detalhes bibliográficos
Autor(a) principal: Sudr?, Adriana Pittella
Data de Publicação: 2012
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFRRJ
Texto Completo: https://tede.ufrrj.br/jspui/handle/jspui/3514
Resumo: Giardia intestinalis infection is important in young dairy goats because it is a wasting protozoosis that compromises animal growth, and there is a risk of environmental contamination by infectious forms that are easily widespread. The aim of this study were: (1) establish the occurrence of Giardia intestinalis in goat fecal samples from two dairy farms (A and B) located in Rio de Janeiro state, Brazil, by fecal parasitological diagnosis; (2) standardize Nested-PCR to improve Giardia sp. detection using ?-giardin, tpi and gdh gene targets; and (3) genotype Giardia intestinalis from positive fecal samples by means of sequencing and genetic analysis, comparing obtained sequences with those retrieved from Genbank. Fecal samples from all young animals (up to one year of age) (n=58) and from 10% of adult animals (n=18) were collected. Samples were collected directly from the rectum and were submitted to centrifuge-flotation in sugar saturate solution for microscopic search of Giardia intestinalis cysts, with further molecular characterization of positive samples. From 76 analyzed samples, 17 (22.4%) were positive for Giardia intestinalis cysts. All positive samples came from young animals of farm A. Also, dog fecal samples (n=9) were collected to evaluate their role in parasite transmission. Three dog samples (33.3%) were positive for Giardia intestinalis cysts. Positive samples were submitted to Nested-PCR and sequencing using ?-giardin, gdh and tpi gene targets. The standardization of molecular methods improved the yeld of DNA fragments, allowing genotyping of 8/17 samples. Only genotype E was observed in isolates from this study, wich presented sequence heterogeneity, with variable numbers of single nucleotide polimorfisms. The present study report the first molecular characterization of Giardia intestinalis from goat samples in Brazil, contributing to a better knowlodge of the molecular epidemiology of this parasite in the country.
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spelling Bomfim, Teresa Cristina Bergamo do609.928.087-15http://lattes.cnpq.br/6299581965425007Figueiredo, Beatriz BrenerCarneiro, Helena L?ciaSantos, Huarrisson AzevedoSilva, Valmir Laurentino091.550.467-70http://lattes.cnpq.br/1820344065898657Sudr?, Adriana Pittella2020-05-12T19:15:13Z2012-12-12Sudr?, Adriana Pittella. Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira. 2012. 108 f. Tese (Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.https://tede.ufrrj.br/jspui/handle/jspui/3514Giardia intestinalis infection is important in young dairy goats because it is a wasting protozoosis that compromises animal growth, and there is a risk of environmental contamination by infectious forms that are easily widespread. The aim of this study were: (1) establish the occurrence of Giardia intestinalis in goat fecal samples from two dairy farms (A and B) located in Rio de Janeiro state, Brazil, by fecal parasitological diagnosis; (2) standardize Nested-PCR to improve Giardia sp. detection using ?-giardin, tpi and gdh gene targets; and (3) genotype Giardia intestinalis from positive fecal samples by means of sequencing and genetic analysis, comparing obtained sequences with those retrieved from Genbank. Fecal samples from all young animals (up to one year of age) (n=58) and from 10% of adult animals (n=18) were collected. Samples were collected directly from the rectum and were submitted to centrifuge-flotation in sugar saturate solution for microscopic search of Giardia intestinalis cysts, with further molecular characterization of positive samples. From 76 analyzed samples, 17 (22.4%) were positive for Giardia intestinalis cysts. All positive samples came from young animals of farm A. Also, dog fecal samples (n=9) were collected to evaluate their role in parasite transmission. Three dog samples (33.3%) were positive for Giardia intestinalis cysts. Positive samples were submitted to Nested-PCR and sequencing using ?-giardin, gdh and tpi gene targets. The standardization of molecular methods improved the yeld of DNA fragments, allowing genotyping of 8/17 samples. Only genotype E was observed in isolates from this study, wich presented sequence heterogeneity, with variable numbers of single nucleotide polimorfisms. The present study report the first molecular characterization of Giardia intestinalis from goat samples in Brazil, contributing to a better knowlodge of the molecular epidemiology of this parasite in the country.A infec??o por Giardia intestinalis assume destaque em caprinos jovens com aptid?o leiteira, pelo fato de ser uma protozoose debilitante, al?m de representar um risco de contamina??o ambiental por formas infectantes, as quais se disseminam facilmente. Este estudo teve como objetivos: (1) estabelecer a ocorr?ncia de Giardia intestinalis em amostras fecais de caprinos, provenientes de duas caprinoculturas (A e B) localizadas no estado do Rio de Janeiro, Brasil, atrav?s do diagn?stico parasitol?gico de fezes; (2) otimizar as melhores condi??es de rea??o da Nested-PCR para detec??o de Giardia sp. utilizando-se os genes alvo ?-giardina, tpi e gdh; e (3) caracterizar o(s) gen?tipo (s) de Giardia intestinalis obtidos de amostras fecais positivas atrav?s de sequenciamento e an?lises gen?ticas, comparando as sequ?ncias obtidas com aquelas depositadas no GenBank. Foram coletadas amostras fecais de todos os animais jovens (at? um ano de idade) (n=58) e de 10% dos animais adultos (n=18). As amostras foram coletadas diretamente do reto e submetidas ? t?cnica de centr?fugo-flutua??o em solu??o saturada de a??car para pesquisa de cistos de Giardia intestinalis por microscopia, com posterior caracteriza??o molecular das amostras positivas. Das 76 amostras analisadas, 17 (22,4%) foram positivas para cistos de Giardia intestinalis, sendo todas provenientes de animais jovens da caprinocultura A. Tamb?m foram coletadas amostras de fezes caninas (n=9) da propriedade A, para avalia??o do papel destes no ciclo de transmiss?o. Tr?s (33,3%) amostras caninas foram positivas para cistos de Giardia. As amostras positivas foram submetidas ?s t?cnicas de PCR-Nested e sequenciamento utilizando-se os genes-alvo ?-giardina, gdh e tpi. A otimiza??o dos m?todos moleculares permitiu uma melhor obten??o de fragmentos de DNA, os quais possibilitaram a genotipagem de 8/17 amostras obtidas. Foi encontrando apenas o gen?tipo E nos isolados, os quais apresentaram heterogeneidade nas sequ?ncias obtidas, com n?meros variados de polimorfismo de nucleot?deo ?nico. O presente estudo foi o primeiro a realizar a caracteriza??o molecular de Giardia intestinalis proveniente de amostras de caprinos no Brasil, contribuindo para tra?ar a epidemiologia molecular deste parasito no pa?s.Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2020-05-12T19:15:12Z No. of bitstreams: 1 2012 - Adriana Pittella Sudre.pdf: 2753081 bytes, checksum: 931fbc50e0213162c92f4472759f8512 (MD5)Made available in DSpace on 2020-05-12T19:15:13Z (GMT). No. of bitstreams: 1 2012 - Adriana Pittella Sudre.pdf: 2753081 bytes, checksum: 931fbc50e0213162c92f4472759f8512 (MD5) Previous issue date: 2012-12-12Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do RJ, FAPERJ, Brasil.application/pdfhttps://tede.ufrrj.br/retrieve/15882/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpghttps://tede.ufrrj.br/retrieve/22178/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpghttps://tede.ufrrj.br/retrieve/28572/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpghttps://tede.ufrrj.br/retrieve/34954/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpghttps://tede.ufrrj.br/retrieve/41414/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpghttps://tede.ufrrj.br/retrieve/47758/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpghttps://tede.ufrrj.br/retrieve/54258/2012%20-%20Adriana%20Pittella%20Sudre.pdf.jpgporUniversidade Federal Rural do Rio de JaneiroPrograma de P?s-Gradua??o em Ci?ncias Veterin?riasUFRRJBrasilInstituto de Veterin?riaABE, N.; READ, C.; THOMPSON, R.C.A; ISEKI, M. Zoonotic genotype of Giardia intestinalis in a Ferret. Journal of Parasitology, v.91, n.1, p. 179-182, 2005. ABE, N.; TERAMOTO, I. 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dc.title.por.fl_str_mv Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
dc.title.alternative.eng.fl_str_mv Standardization of Nested-PCR technique and genotyping of Giardia intestinalis from dairy goats
title Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
spellingShingle Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
Sudr?, Adriana Pittella
Giardia intestinalis
caracteriza??o molecular
caprino
goat
Giardia intestinalis
molecular characterization
Medicina Veterin?ria
title_short Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
title_full Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
title_fullStr Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
title_full_unstemmed Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
title_sort Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira
author Sudr?, Adriana Pittella
author_facet Sudr?, Adriana Pittella
author_role author
dc.contributor.advisor1.fl_str_mv Bomfim, Teresa Cristina Bergamo do
dc.contributor.advisor1ID.fl_str_mv 609.928.087-15
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/6299581965425007
dc.contributor.referee1.fl_str_mv Figueiredo, Beatriz Brener
dc.contributor.referee2.fl_str_mv Carneiro, Helena L?cia
dc.contributor.referee3.fl_str_mv Santos, Huarrisson Azevedo
dc.contributor.referee4.fl_str_mv Silva, Valmir Laurentino
dc.contributor.authorID.fl_str_mv 091.550.467-70
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/1820344065898657
dc.contributor.author.fl_str_mv Sudr?, Adriana Pittella
contributor_str_mv Bomfim, Teresa Cristina Bergamo do
Figueiredo, Beatriz Brener
Carneiro, Helena L?cia
Santos, Huarrisson Azevedo
Silva, Valmir Laurentino
dc.subject.por.fl_str_mv Giardia intestinalis
caracteriza??o molecular
caprino
topic Giardia intestinalis
caracteriza??o molecular
caprino
goat
Giardia intestinalis
molecular characterization
Medicina Veterin?ria
dc.subject.eng.fl_str_mv goat
Giardia intestinalis
molecular characterization
dc.subject.cnpq.fl_str_mv Medicina Veterin?ria
description Giardia intestinalis infection is important in young dairy goats because it is a wasting protozoosis that compromises animal growth, and there is a risk of environmental contamination by infectious forms that are easily widespread. The aim of this study were: (1) establish the occurrence of Giardia intestinalis in goat fecal samples from two dairy farms (A and B) located in Rio de Janeiro state, Brazil, by fecal parasitological diagnosis; (2) standardize Nested-PCR to improve Giardia sp. detection using ?-giardin, tpi and gdh gene targets; and (3) genotype Giardia intestinalis from positive fecal samples by means of sequencing and genetic analysis, comparing obtained sequences with those retrieved from Genbank. Fecal samples from all young animals (up to one year of age) (n=58) and from 10% of adult animals (n=18) were collected. Samples were collected directly from the rectum and were submitted to centrifuge-flotation in sugar saturate solution for microscopic search of Giardia intestinalis cysts, with further molecular characterization of positive samples. From 76 analyzed samples, 17 (22.4%) were positive for Giardia intestinalis cysts. All positive samples came from young animals of farm A. Also, dog fecal samples (n=9) were collected to evaluate their role in parasite transmission. Three dog samples (33.3%) were positive for Giardia intestinalis cysts. Positive samples were submitted to Nested-PCR and sequencing using ?-giardin, gdh and tpi gene targets. The standardization of molecular methods improved the yeld of DNA fragments, allowing genotyping of 8/17 samples. Only genotype E was observed in isolates from this study, wich presented sequence heterogeneity, with variable numbers of single nucleotide polimorfisms. The present study report the first molecular characterization of Giardia intestinalis from goat samples in Brazil, contributing to a better knowlodge of the molecular epidemiology of this parasite in the country.
publishDate 2012
dc.date.issued.fl_str_mv 2012-12-12
dc.date.accessioned.fl_str_mv 2020-05-12T19:15:13Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv Sudr?, Adriana Pittella. Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira. 2012. 108 f. Tese (Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
dc.identifier.uri.fl_str_mv https://tede.ufrrj.br/jspui/handle/jspui/3514
identifier_str_mv Sudr?, Adriana Pittella. Otimiza??o da t?cnica da Nested-PCR e genotipagem de Giardia intestinalis em caprinos de explora??o leiteira. 2012. 108 f. Tese (Programa de P?s-Gradua??o em Ci?ncias Veterin?rias) - Universidade Federal Rural do Rio de Janeiro, Serop?dica.
url https://tede.ufrrj.br/jspui/handle/jspui/3514
dc.language.iso.fl_str_mv por
language por
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